Pinter A, Honnen W J, Tilley S A, Bona C, Zaghouani H, Gorny M K, Zolla-Pazner S
Public Health Research Institute of the City of New York, New York 10016.
J Virol. 1989 Jun;63(6):2674-9. doi: 10.1128/JVI.63.6.2674-2679.1989.
We characterized the structural forms of the human immunodeficiency virus env-encoded proteins with a panel of monoclonal and polyclonal antibodies. Western blot (immunoblot) assays with antibodies specific for gp41 invariably recognized a major component of 160 kilodaltons and a less intense component of 120 kilodaltons in viral lysates. We demonstrated that these species are noncovalently associated tetramers and trimers of gp41 which represent the native form of this protein in virions. These complexes were stable when boiled in the presence of low concentrations of sodium dodecyl sulfate but were dissociated to gp41 monomers at high sodium dodecyl sulfate concentrations. Moreover, two human monoclonal antibodies preferentially recognized the oligomeric complexes over monomeric gp41 in Western blots, indicating the presence of epitopes recognized by the human immune system on the gp41 multimers which are not efficiently expressed by the dissociated monomers. The demonstration of the existence of multimeric env complexes and the enhanced and altered antigenicity of such multimers may be relevant to the design of subunit and recombinant human immunodeficiency virus env vaccines.
我们用一组单克隆和多克隆抗体对人类免疫缺陷病毒env编码蛋白的结构形式进行了表征。用针对gp41的特异性抗体进行的蛋白质印迹（免疫印迹）分析在病毒裂解物中始终识别出一个160千道尔顿的主要成分和一个强度较低的120千道尔顿的成分。我们证明这些物种是gp41的非共价结合四聚体和三聚体，它们代表了该蛋白在病毒颗粒中的天然形式。当在低浓度十二烷基硫酸钠存在下煮沸时，这些复合物是稳定的，但在高浓度十二烷基硫酸钠下会解离成gp41单体。此外，两种人源单克隆抗体在蛋白质印迹中比单体gp41更优先识别寡聚复合物，这表明在gp41多聚体上存在人类免疫系统识别的表位，而解离的单体不能有效表达这些表位。多聚env复合物的存在以及此类多聚体增强和改变的抗原性的证明可能与亚单位和重组人类免疫缺陷病毒env疫苗的设计有关。
